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Separation of transition metals in nonaqueous media with capillary electrophoresis.

The separation of transition metal Ni2+, Cu2+, Co2+, Zn2+, Cd2+ and Fe3+ in methanol was investigated by using different types of organic acids as complexing agents. In pure methanol, the weaker and simpler acetic, propionic, butyric and valeric acids could enhance metal ions selectivity by increasing acid concentration and metal ions could be separated with high efficiency. However, hydroxycarboxylic acids obviously made separation efficiency worse. The effect of mixed organic acids, mixture solvent (methanol-acetonitrile, methanol-water) on metal ions separation was discussed further. The advantages of using nonaqueous solvent over aqueous for metal ions separation were shown finally.

Electrophoresis, Capillary↗

Effect of glucose concentration in the growth medium upon neutral and acidic fermentation end-products of Clostridium bifermentans, Clostridium sporogenes and peptostreptococcus anaerobius.

Clostridium bifermentans, C. sporogenes and Peptostreptococcus anaerobius were grown in Fastidious Anaerobe Broth with glucose concentrations from 0-1.5% (w/v). Gas chromatographic (GLC) analysis of fermentation end-products revealed that a change in glucose concentration markedly affected fermentation patterns of each microorganism. Increased glucose concentration resulted in stimulation of ethanol production by all strains, and of butanol and butyric acid by C. sporogenes. With C. bifermentans, there was a parallel increase in ethanol and acetic acid production and decreased formation of propanol and propionic, iso-butyric and iso-valeric acids. Increased ethanol formation by P. anaerobius was accompanied by an increase in acetic acid and a decrease in butan-1-ol production. The greatest incremental change in GLC profiles occurred when glucose concentration was raised from zero to 0.1% (w/v). These data suggest that glucose concentration in a medium for GLC analysis must be rigorously controlled if reproducible results are to be obtained.

Chromatography, Gas↗

Elucidation of chemical compounds responsible for foot malodour.

Short-chain fatty acids from the socks and feet of subjects either with strong foot odour or with weak or no foot odour were extracted with ethyl ether, and then analysed by gas chromatography/mass spectrometry (GC/MS). Short chain fatty acids were found in greater amounts from those subjects with strong foot odour. Iso-valeric acid was present in all the subjects with foot odour but was not detected in those without. Olfactory evaluations of the various short-chain fatty acid solutions were in agreement with the GC/MS analyses. By incubating sweat and lipid from subjects with strong foot odour, we succeeded in reproducing the foot malodour. GC/MS analyses of reproduced foot odour revealed that short-chain fatty acids were present in a similar composition to that found in vivo.

Adult↗

Acid secretagogue action of structurally gamma-aminobutyric acid (GABA)-related compounds in rats.

The properties of GABA related compounds on gastric function were studied in standardized perfused rat stomach preparations. Intravenous GABA (400 mg/kg) produced a rapid increase in acid secretion. Acid secretagogue actions of 3-aminobutyric acid and 2-aminobutyric acid were less potent than that of GABA. Intravenous injection of 5-amino-n-valeric acid (400 mg/kg) stimulated gastric acid secretion, but 6-amino-n-caproic acid was inactive. Isoguvacine (40 mg/kg, s.c.) stimulated acid output, whereas guvacine, an isomer of isoguvacine, did not. Systemically administered 3-hydroxy-GABA and beta-(p-chlorophenyl)-GABA (PCPGABA) showed significant secretagogue actions. Acid responses to GABA-related compounds were significantly reduced by surgical truncal vagotomy and completely antagonized by atropine. The acid responses to PCPGABA and isoguvacine were partially augmented by yohimbine and propranolol. These results suggest that the secretagogue action of GABA is mimicked by structurally GABA-related compounds, which is mediated through cholinergic receptors, with slight implication of alpha-2 and beta-adrenoceptor mechanisms.

Animals↗

[The effect of monensin on the fermentation of feed with different proportions of hay and concentrate in an artificial rumen (rumen simulation technic)].

An experiment was conducted with rumen pouch (RUSITEC--Rumen Simulation Technique). In four fermentation vessels (V), percent proportions of hay and barley were as follows: V1--40:60, V2 - 60:40, V3--80:20 and V4--100:0. Every day 5 mg of monensin dissolved in 1 ml 96% ethanol were added to each fermentation vessel. All diets were isonitrogenous, and after an addition of urea the crude protein (CP) content made 13% in each diet. The experiment lasted 12 days: so called steady state period took the first six days when the fermentation conditions were stabilized. Monensin reduced dry matter digestibility, production of total volatile fatty acids, acetic acid, n-butyric and isovaleric acids and acetate: propionate proportion, and it increased the production of propionic and n-valeric acids. The production of methane and CO2 decreased. The higher proportion of hay in diets decreased dry matter digestibility, digestibility of detergent fibre, total and individual volatile fatty acids, CO2, methane energy yield of volatile fatty acids (E), glucose utilization, production of adenosine triphosphate and production of fermented hexoses. The production, utilization and recovery of metabolic hydrogen also decreased. The effectiveness of microbial matter synthesis (YATP = 11.3) was highest during the fermentation of feed containing 60% hay and 40% barley.

Animal Feed↗

The effects of isomaltulose, isomalt, and isomaltulose-based oligomers on mineral absorption and retention.

We carried out a balance study to examine the effects of isomaltulose, lactose, isomalt, and isomaltulose-based oligomers (IBOs) on mineral (calcium, magnesium, phosphorus, and iron) absorption and retention. Four-week-old male Wistar rats were divided into five groups of six rats each and fed a basal diet or diet the containing either 5% isomaltulose, 5% lactose, 5% isomalts or isomaltulose-based oligomers (IBOs) ad libitum for 16 d. After 1 wk, the animals were subjected to a 5-d mineral (calcium, magnesium, phosphorus, and iron) balance study. The isomalt feeding, as well as the IBOs feeding, led to significantly elevated mineral absorption and retention. On the other hand, lactose feeding, widely known to enhance calcium absorption, increased only calcium absorption and isomaltulose feeding did not affect mineral absorption or retention. The organic acids in cecum contents were increased by IBOs or isomalt feeding. Succinic and acetic acids in cecum contents were significantly increased by IBOs feeding. Similarly, succinic, acetic, and i-valeric acids and total amount of organic acid in cecum content were significantly increased by isomalt feeding. Although the organic acids in cecum contents were increased by IBOs or isomalt feeding, the pH values and acidity in cecum contents were not changed by IBOs or isomalt feeding. The effect of addition of various organic acids to the mucosal fluid was examined with in vitro study using a hindgut segment. By the addition of acetic acid, and butyric acid, the mineral (calcium, magnesium, and phosphorus) uptake was increased.

Animals↗

VOLATILE FATTY ACID REQUIREMENT OF A STRAIN OF LISTERIA MONOCYTOGENES.

Larson, A. D. (Louisiana State University, Baton Rouge), L. V. Hattier, and C. S. McCleskey. Volatile fatty acid requirement of a strain of Listeria monocytogenes. J. Bacteriol. 89:819-824. 1965.-Listeria monocytogenes strain 2 requires either isobutyric or 2-methylbutyric acid for growth. Elucidation of this requirement began with characterization of the growth-enhancing substance in culture filtrates of Aerobacter aerogenes. A. aerogenes required tryptose, glucose, and aerobic conditions for excretion of active fatty acids into the medium. Commercial preparations of isobutyric, 2-methylbutyric, isovaleric, 3-methylvaleric, and n-valeric acid supported the growth of strain 2. Purification of these fatty acids by gas chromatography demonstrated that strain 2 responded significantly only to isobutyric and 2-methylbutyric acid. Amines, alcohols, and hydroxy fatty acids, which were structurally related to the active acids, did not satisfy the fatty acid requirement of L. monocytogenes strain 2. Only one isolate (strain 2) of 128 cultures of L. monocytogenes required volatile fatty acid for growth.

Alcohols↗

Involvement of spinal NMDA receptors in capsaicin-induced nociception.

Intraplantar injection of capsaicin into the mouse hindpaw produced an acute nociceptive response. The involvement of N-methyl-D-aspartate (NMDA) receptors was examined by intrathecal administration of various excitatory amino acid (EAA) receptor antagonists. The selective and competitive NMDA receptor antagonists, D(-)-2-amino-5-phosphono-valeric acid (APV) and (+/-)-3-(2-carboxypiperazin-4-yl) propyl-1-phosphoric acid (CPP), were most potent in inhibiting the nociceptive response induced by capsaicin (ED50, 0.23 nmol and 0.12 nmol). The noncompetitive NMDA receptor antagonist dizocilpine (MK-801) and the non-NMDA antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) had similar effects on the capsaicin-induced nociception (ED50, 2.90 and 7.98 nmol), while ketamine and 7-chlorokynurenic acid were without effect. Ifenprodil, an antagonist at the receptor-coupled polyamine site, showed a significant reduction of the nociceptive response (ED50, 13.8 nmol). The inhibitory effects of APV, CPP, MK-801, and ifenprodil were reversed by co-administration of NMDA. Coadministration of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) or kainate resulted in a marked reduction of CNQX-induced antinociception. The present results suggest that the NMDA receptor plays a key role in spinal nociceptive processing as measured by the capsaicin test in mice. This nociceptive test may be useful for evaluating competitive NMDA antagonists.

Animals↗

Difference in amounts between titratable acid and total carboxylic acids produced by oral streptococci during sugar metabolism.

The acid produced by the resting cells of Streptococcus mutants NCTC 10449 and HS 6 and S. sanguis ATCC 10556 during sugar metabolism was estimated with a pH-stat and a carboxylic acid analyzer. Lactic, formic, acetic, pyruvic, and carbonic acids were detected in the reaction mixtures, but propionic, citric, succinic, iso-butyric, butyric, iso-valeric, and valeric acids were not detected. The amount of titratable acid estimated by alkaline titration with the pH-stat was larger than the amount of total carboxylic acids estimated with the carboxylic acid analyzer. The difference in quantity between the titratable and the total carboxylic acids increased significantly with an increase in the period of incubation with sugar. Moreover, the value of the alkaline titration of standard lactic, formic, acetic, and pyruvic acids was equal to the amount analyzed with the carboxylic acid analyzer. The results indicated that these two streptococci produced not only these carboxylic acids but also other acid(s), possibly non-carboxylic acid(s), during their sugar metabolism.

Carboxylic Acids↗

Use of volatile fatty acids from an acid-phase digester for denitrification.

This study investigates the ability of naturally-produced volatile fatty acids (VFAs) to act as a carbon source for the removal of nitrate-Nitrogen concentrations in the range of 20 to 200 mg/L. The VFAs were generated from an anaerobic digester treating a 1:1 mixture of starch-rich industrial and municipal wastewater. The experiments were carried out at an ambient temperature of 22+/-2 degrees C using 600 mL batch reactors containing VFA-rich effluent from the digester and prepared primary sludge. Nitrates added in the form of a 1 M potassium nitrate solution were observed to completely disappear in conjunction with significant recovery of pH and alkalinity (i.e. biological denitrification). In all cases, denitrification followed zero-order kinetics with an average rate constant of 2.67 mg/[Lh] and a mean specific denitrification rate of 0.0111 g NOx-N/g volatile suspended solids (VSS) per day. The denitrifier population had a preference for acetic acid which occurred in the largest quantity and was exhausted the most rapidly. The next two most abundant VFAs were butyric acid and propionic acid, which were consumed only after acetate concentrations began to decline. The third "choice" of the denitrifiers was valeric acid, thus it appears that denitrifying bacteria have a sequential preference for VFAs. In addition, they prefer VFAs over other more complex soluble organic carbon forms.

Anaerobiosis↗

Protection by pyroglutamic acid and some of its newly synthesized derivatives against glutamate-induced seizures in mice.

The protection by pyroglutamic acid (CAS 98-79-3) and derivatives Ia-i (injected i.p.) against glutamate- and NMDA (N-methyl-D-aspartate) (i.c.v.) induced seizures in mice has been studied in comparison with known antiepileptics and antagonists of excitatory aminoacids. The potency of pyroglutamic acid and some derivatives (Id,f,g,h) against glutamate-induced convulsions was similar to that shown by glutamic acid diethylester and by valproic acid. Interestingly, pyroglutamic acid did not affect NMDA-induced convulsions which were well antagonized by both 2-amino-5-phosphono valeric acid and by diazepam. Thus, pyroglutamic acid may represent the starting for synthesis of excitatory aminoacid antagonists acting at non NMDA receptors.

Animals↗

Activation of a peroxisome-proliferating catabolite of cholic acid to its CoA ester.

We have shown that a microbial cholic acid catabolite (4R)-4-(2,3,4,6,6a beta,7,8,9,9a alpha,9b beta-decahydro-6a beta-methyl-3-oxo- 1H-cyclopenta[f]quinolin-7 beta-yl)valeric acid (DCQVA), is a potent peroxisome proliferator. In this paper a possible key stage in DCQVA metabolism, the activation of DCQVA to its CoA ester, has been investigated in rat liver microsomes and particulate fractions. The microsomal reaction was dependent on CoA, ATP, DCQVA (0.2-1 mM) and protein content. The reaction was decreased by storage at 4 degrees C, preincubation of microsomes at 37 degrees C for 5 min, or inclusion of Triton X-100 in the reaction mixture. Such treatments also enhanced generation of long-chain fatty acyl-CoAs, as determined by h.p.l.c. analysis. The same effect was caused by exposing the microsomes to phospholipase A2, suggesting that endogenous fatty acids may compete with DCQVA for esterification with CoA. Subcellular fractionation of rat liver demonstrated that the activity of DCQVA-CoA synthesis was localized predominantly in the microsomal fraction, in contrast to long-chain fatty acyl-CoA synthetase, which was distributed among all particulate fractions. Administration of clofibrate of rats did not affect the distribution of DCQVA-CoA synthesis activity. In contrast to a 2-fold induction of long-chain fatty acyl-CoA synthetase by clofibrate treatment, the activity of DCQVA-CoA synthesis in the microsomal fraction decreased by 80%. These results suggest that DCQVA is activated by an enzyme distinct from long-chain fatty acyl-CoA synthetase. The resulting perturbation of fatty acid metabolism may be involved in the mechanism whereby DCQVA causes peroxisome proliferation.

Acyl Coenzyme A↗

The effects of short-chain fatty acids on the neuronal membrane functions of Helix pomatia. I. Electrical properties.

The effects of short-chain fatty acids on the membrane excitability, current-voltage (I-V) characteristics, and cell volume of Helix pomatia neurons were studied. 2-Decenoic acid (DA), having 10 carbon atoms in the hydrocarbon chain, suppressed the excitability of bursting neurons RPa1 (Sakharov and Salanki, 1969) for 30-60 min, while valeric acid (VA), having 5 carbon atoms, had no significant effect on excitability. DA had three different effects on the excitability of beating neurons: in some neurons DA suppressed excitability as in bursting neurons; in a second type of neuron DA had a negligible effect on excitability; and in the neuron located near RPa1 DA had a pentylentetrazol (PTZ)-like effect, i.e., it converted the discharge of the neuron from beating to bursting. DA decreased the peak value of the current, inducing a negative-resistance region in the I-V curve of the bursting neuron without any change in the level of the voltage at which the current reaches its maximal value. DA inhibited the hyperpolarization induced by activation of the Na+ pump, tested after preliminary enrichment of neurons with Na+ ions by incubation in a potassium-free solution for 20 min. DA caused a swelling of the neuron by about 10% which was independent of the Na+ pump. In all the above-mentioned cases VA had no significant effect.

Animals↗

Microbiological degradation of bile acids. Ring A cleavage and 7alpha, 12alpha-dehydroxylation of cholic acid by Arthrobacter simplex.

The metabolism of cholic acid by Arthrobacter simplex was investigated. This organism effected both ring a cleavage and elimination of the hydroxyl groups at C-7 and C-12 and gave a new metabolite, (4R)-4-[4alpha-(2-carboxyethyl)-3aalpha-hexahydro-7abeta-methyl-5-oxoindan-1beta-yl]valeric acid, which was isolated and identified through its partial synthesis. A degradative pathway of cholic acid into this metabolite is tentatively proposed, and the possibility that the proposed pathway could be extended to the cholic acid degradation by other microorganisms besides A. simplex is discussed. The possibility that the observed reactions in vitro could occur during the metabolism of bile acids in vivo is considered.

Arthrobacter↗

Characterization of Clostridium difficile and its differentiation from Clostridium sporogenes by automatic head-space gas chromatography.

Although 47 strains of Clostridium difficile and Clostridium sporogenes were studied by gas chromatography. Acidic and neutral volatile products, formed after 96 h of incubation in glucose-containing peptone yeast-extract medium, were chromatographed. All strains produced appreciable amounts of fatty acids, which were tentatively identified by gas chromatographic retention data. Chromatograms obtained when analysing diethyl ether extracts of culture media of all 47 strains were virtually identical. However, analysis of the broth media by automatic head-space gas chromatography, employing a glass capillary column, gave chromatographic patterns which differentiated the two Clostridium species studied. C. sporogenes was characterized by chromatographic patterns containing a dominant peak of isovaleric acid. Strains of this species produced larger amounts of early eluting compounds and much smaller amounts of butyric and valeric acid than did strains of C. difficile. Automatic head-space gas chromatography provides an efficient means for differentiation of C. difficile and C. sporogenes. This gas chromatographic technique is easier and more rapidly performed than analysis of either extracts of culture media.

Chromatography, Gas↗

N alpha-arylsulfonyl-omega-amidinophenyl-alpha-aminoalkyl-carboxylic acid amides as specific thrombin inhibitors.

Structure-activity relationships for the inhibition of thrombin and trypsin by N alpha-substituted amidinophenyl-alpha-aminoalkylcarboxylic acid amides are presented. Secondary cyclic amides of N alpha-substituted 4-amidinophenylalanine and 2-amino-5-(4-amidinophenyl)valeric acid were found to be potent and specific inhibitors of thrombin, whereas trypsin was inhibited strongly by primary amides of 2-amino-4-(4-amidinophenyl) butyric acid. For this type of inhibitor the carbon amide structure seems to play a decisive role in the enzyme-inhibitor interaction.

Amidines↗

Short chain fatty acids and potassium uptake in a simple lipid-water partition.

Chemical evidence is presented to demonstrate that short chain fatty acids selectively bind potassium instead of sodium in a simple lipid-water partition. The preference for potassium ions as measured by the K+/Na+ relationship increases from valeric acid (C5) to the higher members of the series, reaching a maximum at pelargonic acid (C9) under physiological conditions (37 C and potassium and sodium concentrations as in normal serum). Anions play a very important role in the selection of K+ ions. When NaHCO3, at the concentration present in normal serum, is added to the aqueous phase, a great increase in the absorption of K+ ions is observed, if compared with the same solution without NaHCO3 and at the same pH. A model is described as a working hypothesis which would explain the behavior of the fatty acids.

Blood↗

Hydrolysis and acidification of waste activated sludge at different pHs.

The effect of pH from 4.0 to 11.0 on the hydrolysis and acidification of waste activated sludge (WAS) was investigated. Experimental results showed that at room temperature the sludge hydrolysis was in the following order: alkaline>acidic>(neutral and blank test), and between pH 6.0 and 11.0 the sludge hydrolysis increased with pH. The three main components, soluble protein, carbohydrate and volatile fatty acids (VFAs) in the hydrolytic product were analyzed. It was observed that both the soluble protein and carbohydrate increased with pH in the pH range 7.0-11.0, but also increased to a smaller extent with pH from 7.0 to 4.0. The VFAs concentration was also affected by pH. Under alkaline conditions, the VFAs production was significantly higher than under other conditions. The concentration of VFAs on the 8th day of fermentation at pH 4.0, 7.0 and 10.0 was, respectively, 354.49, 842.00 and 2708.02 mg/L, while VFAs in the blank test was only 633.59 mg/L. The VFAs consisted of acetic, propionic, iso-butyric, n-butyric, iso-valeric and n-valeric acids, but acetic, propionic and iso-valeric were the three main products. Also, the release of soluble phosphorus and ammonia and the production of methane was studied during WAS fermentation at different pHs.

Anaerobiosis↗